Table 2 Staining protocol for autostainer link 48
Step | Category | Reagent | Incubation time (min) |
|---|---|---|---|
1 | Rinse | TBS Automation wash buffer | 0 min |
2 | Endogenous enzyme block | Peroxidazed 1 | 5 min |
3 | Rinse | DI water | 0 min |
4 | Rinse | TBS Automation wash buffer | 5 min |
5 | Primary antibody | Primary antibody placeholder | 30 min |
6 | Rinse | DI water | 0 min |
7 | Rinse | TBS Automation wash buffer | 5 min |
8 | Secondary reagent* | Mouse Linker | 15 min |
9 | Rinse* | DI water | 0 min |
10 | Rinse* | TBS Automation wash buffer | 5 min |
11 | Labelled polymer | HRP polymer | 30 min |
12 | Rinse | DI water | 0 min |
13 | Rinse | TBS Automation wash buffer | 5 min |
14 | Substrate-chromogen | DAB Biocare | 8 min |
15 | Rinse | DI water | 0 min |
16 | Rinse | TBS Automation wash buffer | 5 min |
17 | Enzyme block | Sulfuric acid | 5 min |
18 | Rinse | TBS Automation wash buffer | 0 min |
19 | Rinse | Water | 5 min |
20 | Rinse | TBS Automation wash buffer | 5 min |
21 | Endogenous enzyme block | Peroxidazed 1 | 5 min |
22 | Rinse | DI water | 0 min |
23 | Rinse | TBS Automation wash buffer | 5 min |
24 | Primary antibody | VDAC1 antibody | 30 min |
25 | Rinse | DI water | 0 min |
26 | Rinse | TBS Automation wash buffer | 5 min |
27 | Secondary reagent | Mouse Linker | 15 min |
28 | Rinse | DI water | 0 min |
29 | Rinse | TBS Automation wash buffer | 5 min |
30 | Labelled polymer | HRP polymer | 30 min |
31 | Rinse | DI water | 0 min |
32 | Rinse | TBS Automation wash buffer | 5 min |
33 | Substrate-chromogen | HRP-Magenta | 5 min |
34 | Rinse | DI water | 0 min |
35 | Rinse | TBS Automation wash buffer | 5 min |
36 | Counterstain | Tacha’s Haematoxylin | 3 min |
37 | Rinse | DI water | 0 min |
38 | Rinse | DI water | 5 min |
39 | Rinse | TBS Automation wash buffer | 0 min |